RNeasy 96 QIAcube HT Kit

For automated high-throughput isolation of RNA from animal and human cells and tissue

Simple and reliable automated processing for time and cost savings

Flexible procedures for isolating total RNA or total RNA, including miRNA

Protocols for various starting materials and desired size distribution

High-quality purified RNA ready for use in any downstream application

Consistent RNA yields from small amounts of starting material

The RNeasy 96 QIAcube HT Kit enables simple, automated purification of total RNA or total RNA, including miRNA, from animal and human cells and tissue samples on the QIAcube HT system. Using proven RNeasy silica-membrane technology in a convenient 96-well format, contaminants and inhibitors are removed to yield high-quality RNA ready for downstream analysis.

The RNeasy 96 QIAcube HT Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Automated purification of RNA from tissues.

RNA was isolated from 10 mg of various rat tissues stabilized in RNAlater RNA Stabilization Reagent. RNA was purified using QIAzol Lysis Reagent and the QIAcube HT, BioRobot Universal System, or QIAcube. Purified RNA was analyzed by qRT-PCR for pgk1 mRNA using the QuantiFast Probe RT-PCR Kit. CT values indicate similar performance of RNA extracted with the different purification platforms.

This economical, easy-to-use system provides the same results as other trusted QIAGEN purification technologies (see figure Automated purification of RNA from tissues). Analysis of QIAcube HT purified samples shows high RNA Integrity Scores (RIS) when analyzed with the QIAxcel Advanced system, and performance similar to other automated QIAGEN purification solutions (see figure High-quality RNA from tissues).

* Amounts can vary due to factors such as species, developmental stage, and growth conditions. If the RNeasy procedure enriches for RNA >200 nucleotides, the total RNA yield does not include 5S rRNA, tRNA, and other low molecular weight RNAs, which make up 15–20% of total cellular RNA.

Principle

The RNeasy 96 QIAcube HT Kit enables automated purification of total RNA or total RNA, including miRNA, from 5 x 105 animal or human cells or <40 mg tissue using the QIAcube HT instrument. Six instrument protocols are available, depending on the starting material and desired size distribution of purified RNA. Cell samples can be purified directly using the RNeasy 96 QIAcube HT Kit and the dedicated kit protocol. Tissue samples (<40 mg) and samples for purification of small RNA are processed following sample lysis in QIAzol Lysis Reagent. The procedures yield high-quality RNA that performs well in downstream applications.

The RNeasy 96 QIAcube HT Kit combines the selective binding properties of a silica-based membrane with a high-throughput 96-well format, and is designed for fully-automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument.

Specifications for purification of total RNA from cells

Specification

Description

Number of samples

24–96 samples (to be processed in increments of 8)

Sample input volume

Up to 5 x 105 cells, homogenized in 140 μl Buffer RLT

Elution volume

110 μl

Duration

96 samples in approximately 93 or 114* minutes
24 samples in approximately 54 or 73* minutes

96 samples in approximately 78 minutes
24 samples in approximately 38 minutes

Procedure

Cell lysis is performed manually under highly denaturing conditions with guanidine thiocyanate to immediately inactivate RNases and ensure purification of intact RNA. After the run is started, ethanol is added to the lysate to provide appropriate binding conditions, and the samples are then applied to the wells of the RNeasy 96 plate. Total RNA binds and contaminants are efficiently washed away. High-quality RNA is then eluted in a small volume of RNase-free water, ready for use in any downstream application.

With the automated procedure for purification of total RNA from animal or human cells, all RNA molecules longer than 200 nucleotides are purified. This provides enrichment for mRNA, since most RNAs <200 nucleotides (such as 5.8S rRNA, 5S rRNA, and tRNA, which together comprise 15–20% of total RNA) are selectively excluded. The size distribution of the purified RNA is comparable to that obtained by centrifugation through a CsCl cushion, where small RNA does not sediment efficiently.

For purification of total RNA from tissue, or of total RNA including small RNA, from cells and tissues, we recommend first using QIAzol Lysis Reagent as the lysis buffer. Detailed procedures are provided in the kit handbook.

Applications

The RNeasy 96 QIAcube HT Kit provides efficient, high-throughput RNA sample preparation for research use in fields such as drug screening and basic research. The purified RNA is ready to use in any downstream application, including:

Q Protocol notes for purification of total RNA or total RNA, including miRNA, from animal tissue and cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the miRNeasy 96 QIAcube HT V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

Q Protocol notes for automatic transfer of upper aqueous phase to an empty S-Block on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT pre-treatment V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

Q Protocol notes for purification of total RNA from animal or human cells, including DNase digestion, on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Cell with DNase V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

Q Protocol notes for purification of total RNA from animal or human cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Cell V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

Q Protocol notes for purification of total RNA from tissue, including DNase digestion, on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Tissue with DNase V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

Q Protocol notes for purification of total RNA from tissue on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Tissue V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.

These Q Protocols and installation files can only be used with QIAcube HT Operating software version 4.17 and are not compatible with the new QIAcube HT Prep Manager software.

QIAcube HT Prep Manager Software already includes all standard protocols. Additional protocols and kit configurations can be easily installed. Check for available updates under the "Operating Software" section of the "Product Resources" tab.

Handbook for purification of total RNA, or total RNA, including miRNA, from animal tissue and cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit. For use with QIAcube HT Operating Software 4.17; not compatible with QIAcube HT Prep Manager Software.